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Agdia, Inc (Elkhart, IN) has commercialized a rapid and user-friendly DNA test kit, based on their AmplifyRP® XRT® platform, for detection of Clavibacter michiganensis subsp. michiganensis (Cmm) in tomato.
Cmm, the causal agent of bacterial canker of tomato, is a devastating disease that can result in crop losses of up to 100% if not quickly identified and managed. Agdia’s AmplifyRP XRT test for Cmm provides growers a new and more powerful tool to assist in the diagnosis of this important pathogen.
The test is a real-time isothermal DNA amplification assay that utilizes recombinase polymerase amplification (RPA) technology. RPA facilitates rapid nucleic acid (DNA or RNA) amplification at a single operating temperature of 39°C using crude sample extracts. RPA can provide the same level of sensitivity and specificity as laboratory based PCR methods, but offers some clear advantages. The total assay time is less than 30 minutes, including sample preparation. This is a stark contrast to PCR methods which can take hours or days to complete. PCR analysis must also be completed in a laboratory whereas AmplifyRP XRT can be performed on-site using a battery operated portable fluorometer. Agdia states their new test is capable of detecting concentrations of Cmm bacteria as low as 200 cfu/mL, which offers greater sensitivity than any other commercially available test kit.
To run the test, an end-user first extracts their sample using a mesh extraction bag. After the sample has been extracted a small volume is added to a solution that is used to rehydrate a lyophilized reaction pellet. Once the reaction pellet has been rehydrated, it is added to a portable fluorometer for 20 minutes. During this time Cmm DNA, if present, will be amplified and will produce a fluorescent signal that is detected by the instrument in real-time.
Agdia claims the test is compatible with several commercially available fluorometers but recommends the AmpliFire® manufactured by Douglas Scientific. The AmpliFire is battery operated and has a small footprint, making it possible to test in remote locations. Setup is simple because all the assay parameters are loaded into the AmpliFire using a barcode that is affixed to the tests packaging. It also has a large, intuitive, touch screen interface that is used to initiate reactions and view results. More information on the AmpliFire can be found on Agdia’s website.
Agdia’s AmplifyRP XRT kits for Cmm come with everything needed to perform the test except for the portable fluorometer, pipettes, and tips. The excluded items can be purchased separately. Each kit contains 48 tests. The test is also offered as a service by Agdia Testing Services for those who wish to send samples for diagnosis. For more information on the test contact Agdia at 574-264-2014 or via email at firstname.lastname@example.org
A leading provider of diagnostic solutions for agriculture, Agdia, Inc. has been serving plant breeders, propagators, growers, universities, and private testing laboratories since 1981. The company offers a comprehensive portfolio of validated, easy-to-use diagnostics for identifying plant pathogens, hormones, and transgenic traits, as well as in-house testing services. Agdia’s quality management system is ISO 9001:2008 certified and their Testing Services Laboratory is ISO 17025:2005 accredited.
AmplifyRP XRT Test Kits employ recombinase polymerase amplification (RPA) technology licensed by Agdia, Inc (licensee) from TwistDx Limited, U.K (licensor). Use of the RPA process and probe technologies are protected by US patents 7,270,981 B2, 7,399,590 B2, 7,435,561 B2, 7,485,428 B2 and foreign equivalents in addition to pending patents.
AmplifyRP® is a registered trademark of Agdia, Inc.